PROJECT SUMMARY PAMOC The overarching mission of the WU-DDRCC is to promote collaborative, multidisciplinary research focused on interactions between host and environment in digestive disease. The Precision Animal Models and Organoids Core (PAMOC) evolved over the previous funding period in response to changes in members' needs, and with the establishment of a new institutional gnotobiotic facility. These changes, along with guidance from our External Advisory Board, encouraged us to modify the services and objectives of this Core. Accordingly, our current objectives are to 1) assist investigators in the efficient and cost-effective development and propagation of genetically altered mouse models to elucidate the pathogenesis of digestive diseases; and 2) assist investigators in the culture of human and mouse gastrointestinal epithelial cell organoids to address cell biological questions relevant to digestive diseases and host-environment interactions. Both human and mouse gastrointestinal epithelial cells and the required media are available to DDRCC investigators for growth and differentiation of organoids. These novel models will serve as important avenues for pre-clinical studies and enable translation of basic research advances into clinical practice. KEY ACCOMPLISHMENTS since 2013 include: completed 24 nucleic acid injection projects (both conventional DNA transgenes and CRISPR/Cas9/targeting vector injections) for 7 members; 3 embryonic stem cell injection projects for 2 members; and 11 withdrawals of transgenic/knockin mice from the Mouse Bank by 9 members. Gnotobiotic services (provision of germ-free mice, rederivation to germ-free status, and gnotobiotic conventionalization) were provided to 6 members. Related to the proposed Organoid component, we have already distributed human cell lines as well as wild type and reporter mouse cell lines to 13 members. We also provided L-WRN conditioned media to 9 members. Since 2013, 22 members used the Core, resulting in 57 publications citing the WU-DDRCC. Our SPECIFIC AIMS include: (1) To assist investigators in the generation and propagation of transgenic, knockout, and knockin mice for the study of digestive diseases. This includes injections of nucleic acids, including CRISPR/Cas9 and TALEN reagents, into zygotes, assistance with rederivation of lines as specific pathogen free, and in vitro fertilization. (2) To maintain a ?Mouse Bank? containing transgenic strains of use to DDRCC investigators and to provide these mice on demand for digestive disease-relevant projects. (3) To provide banked human and mouse gastrointestinal epithelial cell lines and the culture media required for successful growth of organoids and for growth of epithelial cell monolayers. Support for monitoring cell growth and for imaging organoids will also be provided. (4) To provide assistance with the derivation of new gastrointestinal epithelial cell lines from mice and humans. Aims 1 and 2 will continue to leverage the outstanding facilities, personnel, and expertise of the Washington University Institutional Mouse Genetics Core. Aims 3 and 4 will benefit from infrastructure and expertise developed by Dr. Ciorba and his collaborators here at Washington University.